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Breeding method of strong-acid resistant lactic acid bacteria strain

A technology of acid lactic acid bacteria and lactic acid bacteria, applied in the field of microbial genetic breeding and fermentation engineering, can solve the problems of limited high-quality probiotics and rare applications of microorganisms, and achieve the effect of genetic stability

Inactive Publication Date: 2016-09-07
SUZHOU JIANSHIXING BIOLOGICAL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Radiation mutagenesis is a kind of physical mutagenesis. Mutagens mainly include ultraviolet rays, X-rays, γ-rays, and fast neutrons. very limited
There is another type of mutagenesis—chemical mutagenesis. Mutagens are mainly base analogs, alkylating agents, etc., which are widely used in plants, but rarely used in microorganisms.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] (1) Preparation of chemical mutagen solution

[0031] First dissolve a certain amount of nitrosoguanidine with a small amount of dimethyl sulfoxide (DMSO), and then prepare a 1.0 g / L mother solution with 0.1 M sterile phosphate buffer (pH 7.0).

[0032] (2) Preparation of artificial gastric juice

[0033] Dissolve 0.2g NaCl and 0.35g pepsin in an appropriate amount of distilled water, adjust the pH value to 2.0 with HCl, and set the volume to 100ml; then, filter through a 0.22µm microporous membrane for use.

[0034] (3) Culture medium preparation

[0035] MRS liquid medium: 10g / L peptone, 10g / L beef extract, 5g / L yeast powder, 20g / L glucose, 5g / L anhydrous sodium acetate, 1ml / L Tween-80, 2g / L diamine citrate , 2g / L dipotassium hydrogen phosphate, 0.58g / L magnesium sulfate, 0.25g / L manganese sulfate, dissolved in distilled water, adjusted to pH 6.6-6.8, and constant volume; high pressure (1×10 5 Pa) Sterilize for 20min.

[0036] MRS plate medium: Add 20.0g / L agar on t...

Embodiment 2

[0047] (1) Preparation of chemical mutagen solution

[0048] First dissolve a certain amount of nitrosoguanidine with a small amount of dimethyl sulfoxide (DMSO), and then prepare a 1.0 g / L mother solution with 0.1 M sterile phosphate buffer (pH 7.0).

[0049] (2) Preparation of artificial gastric juice

[0050] Dissolve 0.2g NaCl and 0.35g pepsin in an appropriate amount of distilled water, adjust the pH value to 2.0 with HCl, and set the volume to 100ml; then, filter through a 0.22µm microporous membrane for use.

[0051] (3) Culture medium preparation

[0052] MRS liquid medium: 10g / L peptone, 10g / L beef extract, 5g / L yeast powder, 20g / L glucose, 5g / L anhydrous sodium acetate, 1ml / L Tween-80, 2g / L diamine citrate , 2g / L dipotassium hydrogen phosphate, 0.58g / L magnesium sulfate, 0.25g / L manganese sulfate, dissolved in distilled water, adjusted to pH 6.6-6.8, and constant volume; high pressure (1×10 5 Pa) Sterilize for 20min.

[0053] MRS plate medium: Add 20.0g / L agar on...

Embodiment 3

[0064] (1) Preparation of chemical mutagen solution

[0065] First dissolve a certain amount of nitrosoguanidine with a small amount of dimethyl sulfoxide (DMSO), and then prepare a 1.0 g / L mother solution with 0.1 M sterile phosphate buffer (pH 7.0).

[0066] (2) Preparation of artificial gastric juice

[0067] Dissolve 0.2g NaCl and 0.35g pepsin in an appropriate amount of distilled water, adjust the pH value to 2.0 with HCl, and set the volume to 100ml; then, filter through a 0.22µm microporous membrane for use.

[0068] (3) Culture medium preparation

[0069] MRS liquid medium: 10g / L peptone, 10g / L beef extract, 5g / L yeast powder, 20g / L glucose, 5g / L anhydrous sodium acetate, 1ml / L Tween-80, 2g / L diamine citrate , 2g / L dipotassium hydrogen phosphate, 0.58g / L magnesium sulfate, 0.25g / L manganese sulfate, dissolved in distilled water, adjusted to pH 6.6-6.8, and constant volume; high pressure (1×10 5 Pa) Sterilize for 20min.

[0070] MRS plate medium: Add 20.0g / L agar on...

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Abstract

The invention discloses a breeding method of strong-acid resistant lactic acid bacteria strain. The breeding method includes inducing common probiotic lactic acid bacteria by chemical mutagen such as nitrosoguanidine, exploring massive mutation colony of lactic acid as a new lactic acid genetic germplasm resource library, screening mutant strains, which are capable of growing on lactobacillus culture medium MRS and have regular shape and distinctive characteristics, from the resource library, and screening strains that could multiply fast after being treated by simulated gastric fluid from the mutant strains, which are the novel high-quality and strong-acid resistant lactic acid bacteria strain.

Description

technical field [0001] The invention relates to a breeding method for strong-acid-resistant lactic acid bacteria strains, in particular to developing mutant populations and breeding new high-quality probiotic lactic acid bacteria through chemical mutagenesis, and belongs to the technical field of microbial genetic breeding and fermentation engineering. Background technique [0002] There are a variety of flora in the digestive system of the human gastrointestinal tract, including beneficial, harmful and neutral. Under normal circumstances, there is a natural balance of these flora. However, under the modern diet, high pressure of life and work, and pathological conditions, the number of harmful flora increases, and the neutral flora produces harmful substances to the human body, turning into harmful flora, which leads to the natural balance of the human colon and digestive system flora Out of balance, susceptible to infection by pathogenic bacteria, the human body is in a su...

Claims

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Application Information

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IPC IPC(8): C12N15/01C12N1/20
CPCC12N1/20C12N15/01
Inventor 刘世名
Owner SUZHOU JIANSHIXING BIOLOGICAL SCI & TECH CO LTD
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